Des(rhamnosyl)verbascoside抗HBV作用的定量蛋白质组学研究

为探究des(rhamnosyl)verbascoside体外抗乙型肝炎病毒(HBV)的活性作用和机制,本研究以des(rhamnosyl)verbascoside为实验药物对HepG2.2.15细胞进行干预,实验分为药物干预组和对照组,采用串联质谱标签(Tandem Mass Tag,TMT)蛋白质组学方法对提取的总蛋白进行分析。结果表明,共筛选得到300个差异表达蛋白,其中有109个上调蛋白,191个下调蛋白。基因本体论(Gene Ontology,GO)分析结果显示,差异蛋白主要参与DNA复制(DNA replication)、鞘糖脂代谢(Glycosphingolipid metabolic process)、细胞增殖(Cell proliferation)、寡糖分解代谢(Oligosaccharide catabolic process)等生物学过程,以及DNA聚合酶活性(DNA polymerase activity)、丝氨酸型羧肽酶活性(Serine type carboxypeptidase activity)、DNA引物酶活性(DNA primase activity)等分子功能。京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析结果显示,差异蛋白主要参与细胞代谢(Metabolism)、遗传信息传导(Genetic information processing)、生物系统通路(Organismal systems)等相关信号通路。亚细胞定位分析表明,差异蛋白大多定位在细胞质和细胞核。本研究共筛选出13个与抗HBV密切相关的蛋白。通过定量蛋白组学初步揭示des(rhamnosyl)verbascoside可能通过增加HGF、SORT1、MAN2B1,减少PRIM1、PRIM2、POLA1、POLD3、POLD2、POLD1、POLE、ERCC2、LAMC1、SDC1等蛋白表达来起到体外抗HBV的作用。

Verbascoside exerts an anti-atherosclerotic effect by regulating liver glycerophospholipid metabolism

Verbascoside,abundant in olive mill wastewater,is a phenylethanolic glycoside with a wide range of pharmacological activities.Atherosclerosis(AS)is a common metabolic disease and abnormal lipid metabolism in liver is inseparable from its formation and development.In this study,the anti-atherosclerotic effect of verbascoside was evaluated by establishing an atherosclerosis model based on western diet feeding of apolipoprotein E-defi cient mice for 16 weeks.After 12 weeks of administration during the feeding period,the levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)in the plasma of mice were signifi cantly decreased,the formation of arterial plaques was delayed,and the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and lactate dehydrogenase(LDH)in plasma were alleviated,showing the hepatoprotective effect.In addition,based on untargeted lipidomic analysis,verbascoside stabilized glycerophospholipid metabolism,modulated lipid metabolism disorders and reduced lipid deposition in the liver to achieve the therapeutic effi cacy against atherosclerosis by regulating cardiolipin(CL),ether-linked phosphatidylcholine(ether-PC),lysophophatidylcholine(LPC),phosphatidylcholine(PC),oxidized phosphatidylcholine(OxPC),oxidized phosphatidylethanolamine(OxPE),triacylglycerol(TG),sphingomyelin(SM)back to normal levels.

Mechanism of action of verbascoside in mice with Parkinson's disease based on molecular docking,molecular dynamics and in vivo experiments

Objective:To explore the improving effect of verbascoside on Parkinson's disease(PD)model mice and to clarify the possible mechanism.Methods:60 C57BL/6 male mice were randomly divided into normal group,model group,low,middle and high dose groups(30,60 and 120 mg/kg,respectively).After a week of adaptive culture,except for the blank group,the subacute model of PD was established by intraperitoneal injection of 1-methyl-4-phenyl-1(MPTP)in each group for 5 consecutive days.After a week of adaptive culture,except for the blank group,the subacute model of PD was established by intraperitoneal injection of 1-methyl-4-phenyl-1(MPTP)in each group for 5 consecutive days.The establishment of the model,the drug group was intragastrically administered continuously for 7 d,and the mice in the normal group were injected with the same amount of saline.The behavioral changes of mice were observed by open field test,pole climbing test and grip test;the morphology,apoptosis and Nissl bodies of substantia nigra neurons were observed by HE staining and Nissl staining;the ultrastructural changes of neurons were observed by transmission electron microscope;the positive expressions of tyrosine hydroxylase(TH),α-synuclein(α-Syn)and toll-like receptor 4(TLR4)were detected by immunohistochemistry.The protein levels of TH,α-Syn,TLR4,NF-κB and P-NF-κB in substantia nigra of mice were detected by Western blot,and the levels of inflammatory factors IL-1β,IL-6 and tumor necrosis factor-α(TNF-α)in serum were detected by Elisa.Molecular docking was used to verify the binding ability of verbascoside to TLR4/NF-κB signal pathway and related factors,and molecular dynamics was used to verify the binding mode and stability of verbascoside with compounds.Results:compared with the normal group,the total walking distance,average speed,free activity time,pole climbing time and forelimb grip in the model group were significantly longer than those in the normal group(P<0.05).The ultrastructural changes of neurons,nuclear lysis,deformation and mitochondrial damage were observed under transmission electron microscope,and the number and morphological changes of HE staining substantia nigra neurons were decreased(P<0.05).Nissl staining showed that the number of Nissl positive cells decreased(P<0.05),the positive expression of TH decreased and the positive expression of α-syn and TLR4 increased(P<0.05).Western blotting showed that the expression of TH protein decreased,the expression of α-syn,TLR4,NF-κB p65 protein increased(P<0.05),and the expression of IL-1β,IL-6 and TNF-α in serum increased significantly(P<0.05).Compared with the model group,the activity ability of mice in the treatment group was significantly improved(P<0.05),the morphology of neurons gradually recovered and the number of neurons increased(P<0.05).The number of Nissl positive cells increased(P<0.05).The expression of TH protein increased,while the expression of α-syn,TLR4,NF-κBp65 protein decreased(P<0.05),and the expression of IL-1β,IL-6,TNF-α in serum decreased significantly(P<0.05).The results of molecular docking and molecular dynamics simulation show that the combination of mulberry glycoside with the compound is good and the property is stable.Conclusion:verbascoside has protective effect on PD mice,and its mechanism may be related to TLR4/NF-κB pathway,and then regulate neuroimmunity.

Verbascoside promotes the regeneration of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra

Tyrosine hydroxylase is a key enzyme in dopamine biosynthesis. Change in tyrosine hydroxylase expression in the nigrostriatal system is closely related to the occurrence and development of Parkinson＇s disease. Verbascoside, an extract from Radix Rehmanniae Praeparata has been shown to be clinically effective in treating Parkinson＇s disease. However, the underlying mechanisms remain unclear. It is hypothesized that the effects of verbascoside on Parkinson＇s disease are related to tyrosine hydroxylase expression change in the nigrostriatal system. Rat models of Parkinson＇s disease were established and verbascoside（60 mg/kg） was administered intraperitoneally once a day. After 6 weeks of verbascoside treatment, rat rotational behavior was alleviated; tyrosine hydroxylase m RNA and protein expression and the number of tyrosine hydroxylase-immunoreactive neurons in the rat right substantia nigra were significantly higher than the Parkinson＇s model group. These findings suggest that the mechanism by which verbascoside treats Parkinson＇s disease is related to the regeneration of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra.

Verbascoside—A Review of Its Antitumor Activities

Cancer is a set of diseases including abnormal growth of cells that can spread to another tissue. Verbascoside (or acteoside) is a naturally occurring, water-soluble secondary metabolite with significant biological properties, which is distributed widely in plant kingdom. Verbascoside is pharmacologically active compounds with many recent evidences that support its biological activities and safety. This review focuses on the recent studies that concerned with the antitumor activities of verbascoside alone and as a synergistic agent as well as nanoproduct. It also shows the latest advances in its antitumor effects, cytotoxic selectivity and its efficiencies in treating cancer, in vitro and/or vivo.

Complete biosynthesis of the phenylethanoid glycoside verbascoside

Verbascoside,which was first discovered in 1963,is a well-known phenylethanoid glycoside(PhG)that exhibits antioxidant,anti-inflammatory,antimicrobial,and neuroprotective activities and contributes to the therapeutic effects of many medicinal plants.However,the biosynthetic pathway of verbascoside remains to be fully elucidated.Here,we report the identification of two missing enzymes in the verbascoside biosynthesis pathway by transcriptome mining and in vitro enzymatic assays.Specifically,a BAHD acyltransferase(hydroxycinnamoyl-CoA:salidroside hydroxycinnamoyltransferase[SHCT])was shown to catalyze the regioselective acylation of salidroside to form osmanthuside A,and a CYP98 hydroxylase(osmanthuside B 3,30-hydroxylase[OBH])was shown to catalyze meta-hydroxylations of the p-coumaroyl and tyrosol moieties of osmanthuside B to complete the biosynthesis of verbascoside.Because SHCTs and OBHs are found in many Lamiales species that produce verbascoside,this pathway may be general.The findings from the study provide novel insights into the formation of caffeoyl and hydroxytyrosol moieties in natural product biosynthetic pathways.In addition,with the newly acquired enzymes,we achieved heterologous production of osmanthuside B,verbascoside,and ligupurpuroside B in Escherichia coli;this work lays a foundation for sustainable production of verbascoside and other PhGs in micro-organisms.

Efficacy of verbascoside,echinacoside,crenatoside on altitudeinduced fatigue in rats and possible mechanism

OBJECTIVE:To study the efficacy and mechanism of three phenylethanoid glycosides(PhG s)(verbascoside,echinacoside,and crenatoside)on altitude-induced fatigue in rats.METHODS:Altitude-induced fatigue model rats were established in a large hypobaric chamber.Swimming time,energy storage substances,metabolic enzymes,and metabolites were used to evaluate the anti-fatigue activities and mechanism of three PhG s(verbascoside,echinacoside,and crenatoside)(150 mg/kg,intragastric administration)in the hypoxic environment.RESULTS:The three PhGs,especially verbascoside,could prolong the swimming time of rats,ameliorate the edema and inflammatory infiltration of liver and skeletal muscle,increase the level of energy storage substances,reduce the decomposition of proteins,and exhibit positive effects on the metabolism-related enzyme activity and metabolites.CONCLUSIONS:The PhG s,especially verbascoside,are very potential with anti-fatigue activity in hypoxia.The mechanism may be explained with regulation of energy metabolism and reduction of oxidative stress.

Fast repair of dAMP hydroxyl radical adduct by verbascoside via electron transfer

DNA damaged by oxygen radicals has been implicated as a causative event in a number of degenerative diseases, including cancer and aging. So it is very impotant to look for ways in which either oxygen radicals are scavenged prior to DNA damage or damaged DNA is repaired to supplement the cells' inadequate repair capacity. The repair activity and its mechanism of verbascoside, isolated from Pedicularis species, towards dAMP-OH· was studied with pulse radiolytic technique. On pulse irradiation of nitrous oxide saturated 2 mmol/L dAMP aqueous solution containing verbascoside, the transient absorption spectrum of the hydroxyl adduct of dAMP decayed with the formation of that of the phenoxyl radical of verbascoside well under 100 microseconds after electron pulse irradiation. The result indicated that dAMP hydroxyl adducts can be repaired by verbascoside. The rate constants of the repair reaction was deduced to be 5.9 x 108 dm3 · mol-1 · s-1 . A deeper understanding of this new repair mechanism will undoubtedly help researchers explore new prevent and/or intervening medicine.

PROTECTIVE EFFECTS OF VERBASCOSIDE ON STATIN-INDUCED MYOTOXICITY IN ZEBRAFISH EMBRYOS

Myotoxicity is a well-known side effect of statins,the common lipid-lowering drug.Herba Cistanches(HC,Cistanche deserticola)is a Chinese herb traditionally used for muscle problems.Recent studies demonstrated that HC could reduce muscle damage in post-exercised rats.Verbascoside(Vb)was fournd as the major constituent in the active fraction of HC and is known to be muscle protective,

Antifungal activity of total and fractionated phenolic extracts from two wild edible herbs

The chemical characterization of phenolic extracts of salad burnet (Sanguisorba minor Scop. s.l.) and broomrape (Orobanche crenata Forsk.) was performed. Active fractions were separated and characterized and, in the case of O. crenata, two phenolic compounds, verbascoside and is overbascoside, were quantitatively extracted, purified and characterized. These compounds proved to be effective in reducing the growth of some important fungi responsible for the decay of fresh fruit and vegetables in postharvest. Verbascoside showed significant activity against Penicillium italicum, fairly inhibited Monilinia laxa, P. expansum, and Aspergillus carbonarius, and showed little activity against Botrytis cinerea, M. fructicola and P. digitatum. Isoverbascoside completely inhibited B. cinerea, P. digitatum, P. italicum, and P. expansum;it fairly inhibited M. laxa and A. carbonarius;while it showed to be less effective against M. fructicola. The obtained results support the possible use of some wild edible plants as a source of phenolic substances for the postharvest biological control of fresh fruit and vegetable rots.

苯丙素苷化合物的合成研究进展

生物活性有机化合物的合成是有机化学的重要前沿领域之一。苯丙素苷是一类天然糖苷类化合物的总称,具有抗肿瘤、抗病毒、抗氧化、消炎等生物活性,其全合成是一项具有重要科学意义和挑战性的研究工作。本论文对具有广泛生物活性的天然化合物糖苷苯丙素苷全合成研究进展进行了概述。